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11: Peptide Neutralization
Blocking (neutralizing) peptides are available as negative controls for all Santa Cruz Biotechnology, Inc. affinity-purified rabbit and goat polyclonal antibodies and monoclonal antibodies raised against peptide antigens. Antibody binding to antigen may be blocked/competed by pre-absorption with the blocking peptide.
Determine the highest antibody dilution at which a consistently positive result is achieved for the desired test. For example, H-Ras (259) is recommended for immunoprecipitation at 1 µg/ml but is positive at a dilution of 50 ng/ml.
For blocking/competition, combine antibody (at a concentration determined by the aforementioned method) with a five-fold (by weight) excess of blocking peptide in a small volume (500 µl) of PBS (Buffers and General Solutions). Incubate for up to 2 hours at room temperature or overnight at 4º C.
Following blocking/competition, dilute antibody/peptide mixture into appropriate blocking buffer and proceed with the desired research application.
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